The expression and secretion of human lactoferrin in Bacillus subtilis].

Yuting Zhang, Yang Li, Yaokang Wu, Yanfeng Liu, Jianghua Li, Guocheng DU, Xueqin Lv, Long Liu

Sheng wu gong cheng xue bao = Chinese journal of biotechnology 2024 Jun 25

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Human lactoferrin (HLF), an essential nutrient found in breast milk, possesses antibacterial, anti-inflammatory, and immune-enhancing properties. In this study, the effects of three constitutive promoters (P21, P43, and Pveg) and three inducible promoters (Pgrac100, PxylA, and Ptet*) on the expression of HLF were compared using Bacillus subtilis G601 as the host strain. The results showed that the highest expression of HLF, reaching 651.57 μg/L, was achieved when regulated by the Ptet* promoter. Furthermore, the combinational optimization of ribosome binding site (RBS) and signal peptides was investigated, and the optimal combination of RBS6 and SPyycP resulted in increased HLF expression to 1 099.87 μg/L, with 498.68 μg/L being secreted extracellularly. To further enhance HLF secretion, the metal cations-related gene dltD was knocked out, leading to an extracellular HLF level of 637.28 μg/L. This study successfully demonstrated the secretory expression of HLF in B. subtilis through the selection and optimization of expression elements, laying the foundation for the development of efficient B. subtilis cell factories for lactoprotein synthesis.

Citation

Yuting Zhang, Yang Li, Yaokang Wu, Yanfeng Liu, Jianghua Li, Guocheng DU, Xueqin Lv, Long Liu. The expression and secretion of human lactoferrin in Bacillus subtilis]. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2024 Jun 25;40(6):1895-1908